Cargando…

The gene expression profiles of induced pluripotent stem cells from individuals with childhood cerebral adrenoleukodystrophy are consistent with proposed mechanisms of pathogenesis

INTRODUCTION: X-linked adrenoleukodystrophy (X-ALD) is a complex disorder with variable expressivity that affects the nervous, adrenocortical and male reproductive systems. Although ABCD1 mutations are known to provide the genetic basis for X-ALD, its pathogenesis is not fully elucidated. While elev...

Descripción completa

Detalles Bibliográficos
Autores principales: Wang, Xiao-Ming, Yik, Wing Yan, Zhang, Peilin, Lu, Wange, Dranchak, Patricia K, Shibata, Darryl, Steinberg, Steven J, Hacia, Joseph G
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3580430/
https://www.ncbi.nlm.nih.gov/pubmed/23036268
http://dx.doi.org/10.1186/scrt130
_version_ 1782260245685338112
author Wang, Xiao-Ming
Yik, Wing Yan
Zhang, Peilin
Lu, Wange
Dranchak, Patricia K
Shibata, Darryl
Steinberg, Steven J
Hacia, Joseph G
author_facet Wang, Xiao-Ming
Yik, Wing Yan
Zhang, Peilin
Lu, Wange
Dranchak, Patricia K
Shibata, Darryl
Steinberg, Steven J
Hacia, Joseph G
author_sort Wang, Xiao-Ming
collection PubMed
description INTRODUCTION: X-linked adrenoleukodystrophy (X-ALD) is a complex disorder with variable expressivity that affects the nervous, adrenocortical and male reproductive systems. Although ABCD1 mutations are known to provide the genetic basis for X-ALD, its pathogenesis is not fully elucidated. While elevated very long chain fatty acid (VLCFA) levels in blood and reduced VLCFA catabolic activity in cultured fibroblasts are biomarkers used to identify ABCD1 mutation carriers, the roles peroxisomal lipid metabolism play in disease etiology are unknown. METHODS: Primary skin fibroblasts from two male patients with the childhood cerebral form of the disease (CCALD) caused by ABCD1 frameshift or missense mutations and three healthy donors were transduced with retroviral vectors expressing the OCT4, SOX2, KLF4 and c-MYC factors. Candidate induced pluripotent stem cells (iPSCs) were subject to global gene expression, DNA methylation, DNA copy number variation, and genotyping analysis and tested for pluripotency through in vitro differentiation and teratoma formation. Saturated VLCFA (sVLCFA) and plasmalogen levels in primary fibroblasts and iPSCs from healthy donors as well as CCALD patients were determined through mass spectroscopy. RESULTS: Skin fibroblasts from CCALD patients and healthy donors were reprogrammed into validated iPSCs. Unlike fibroblasts, CCALD patient iPSCs show differentially expressed genes (DEGs) relevant to both peroxisome abundance and neuroinflammation. Also, in contrast to fibroblasts, iPSCs from patients showed no significant difference in sVLCFA levels relative to those from controls. In all cell types, the plasmalogen levels tested did not correlate with ABCD1 mutation status. CONCLUSION: Normal ABCD1 gene function is not required for reprogramming skin fibroblasts into iPSCs or maintaining pluripotency. Relative to DEGs found in fibroblasts, DEGs uncovered in comparisons of CCALD patient and control iPSCs are more consistent with major hypotheses regarding disease pathogenesis. These DEGs were independent of differences in sVLCFA levels, which did not vary according to ABCD1 mutation status. The highlighted genes provide new leads for pathogenic mechanisms that can be explored in animal models and human tissue specimens. We suggest that these iPSC resources will have applications that include assisting efforts to identify genetic and environmental modifiers and screening for therapeutic interventions tailored towards affected cell populations and patient genotypes.
format Online
Article
Text
id pubmed-3580430
institution National Center for Biotechnology Information
language English
publishDate 2012
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-35804302013-02-26 The gene expression profiles of induced pluripotent stem cells from individuals with childhood cerebral adrenoleukodystrophy are consistent with proposed mechanisms of pathogenesis Wang, Xiao-Ming Yik, Wing Yan Zhang, Peilin Lu, Wange Dranchak, Patricia K Shibata, Darryl Steinberg, Steven J Hacia, Joseph G Stem Cell Res Ther Research INTRODUCTION: X-linked adrenoleukodystrophy (X-ALD) is a complex disorder with variable expressivity that affects the nervous, adrenocortical and male reproductive systems. Although ABCD1 mutations are known to provide the genetic basis for X-ALD, its pathogenesis is not fully elucidated. While elevated very long chain fatty acid (VLCFA) levels in blood and reduced VLCFA catabolic activity in cultured fibroblasts are biomarkers used to identify ABCD1 mutation carriers, the roles peroxisomal lipid metabolism play in disease etiology are unknown. METHODS: Primary skin fibroblasts from two male patients with the childhood cerebral form of the disease (CCALD) caused by ABCD1 frameshift or missense mutations and three healthy donors were transduced with retroviral vectors expressing the OCT4, SOX2, KLF4 and c-MYC factors. Candidate induced pluripotent stem cells (iPSCs) were subject to global gene expression, DNA methylation, DNA copy number variation, and genotyping analysis and tested for pluripotency through in vitro differentiation and teratoma formation. Saturated VLCFA (sVLCFA) and plasmalogen levels in primary fibroblasts and iPSCs from healthy donors as well as CCALD patients were determined through mass spectroscopy. RESULTS: Skin fibroblasts from CCALD patients and healthy donors were reprogrammed into validated iPSCs. Unlike fibroblasts, CCALD patient iPSCs show differentially expressed genes (DEGs) relevant to both peroxisome abundance and neuroinflammation. Also, in contrast to fibroblasts, iPSCs from patients showed no significant difference in sVLCFA levels relative to those from controls. In all cell types, the plasmalogen levels tested did not correlate with ABCD1 mutation status. CONCLUSION: Normal ABCD1 gene function is not required for reprogramming skin fibroblasts into iPSCs or maintaining pluripotency. Relative to DEGs found in fibroblasts, DEGs uncovered in comparisons of CCALD patient and control iPSCs are more consistent with major hypotheses regarding disease pathogenesis. These DEGs were independent of differences in sVLCFA levels, which did not vary according to ABCD1 mutation status. The highlighted genes provide new leads for pathogenic mechanisms that can be explored in animal models and human tissue specimens. We suggest that these iPSC resources will have applications that include assisting efforts to identify genetic and environmental modifiers and screening for therapeutic interventions tailored towards affected cell populations and patient genotypes. BioMed Central 2012-10-04 /pmc/articles/PMC3580430/ /pubmed/23036268 http://dx.doi.org/10.1186/scrt130 Text en Copyright ©2012 Wang et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Wang, Xiao-Ming
Yik, Wing Yan
Zhang, Peilin
Lu, Wange
Dranchak, Patricia K
Shibata, Darryl
Steinberg, Steven J
Hacia, Joseph G
The gene expression profiles of induced pluripotent stem cells from individuals with childhood cerebral adrenoleukodystrophy are consistent with proposed mechanisms of pathogenesis
title The gene expression profiles of induced pluripotent stem cells from individuals with childhood cerebral adrenoleukodystrophy are consistent with proposed mechanisms of pathogenesis
title_full The gene expression profiles of induced pluripotent stem cells from individuals with childhood cerebral adrenoleukodystrophy are consistent with proposed mechanisms of pathogenesis
title_fullStr The gene expression profiles of induced pluripotent stem cells from individuals with childhood cerebral adrenoleukodystrophy are consistent with proposed mechanisms of pathogenesis
title_full_unstemmed The gene expression profiles of induced pluripotent stem cells from individuals with childhood cerebral adrenoleukodystrophy are consistent with proposed mechanisms of pathogenesis
title_short The gene expression profiles of induced pluripotent stem cells from individuals with childhood cerebral adrenoleukodystrophy are consistent with proposed mechanisms of pathogenesis
title_sort gene expression profiles of induced pluripotent stem cells from individuals with childhood cerebral adrenoleukodystrophy are consistent with proposed mechanisms of pathogenesis
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3580430/
https://www.ncbi.nlm.nih.gov/pubmed/23036268
http://dx.doi.org/10.1186/scrt130
work_keys_str_mv AT wangxiaoming thegeneexpressionprofilesofinducedpluripotentstemcellsfromindividualswithchildhoodcerebraladrenoleukodystrophyareconsistentwithproposedmechanismsofpathogenesis
AT yikwingyan thegeneexpressionprofilesofinducedpluripotentstemcellsfromindividualswithchildhoodcerebraladrenoleukodystrophyareconsistentwithproposedmechanismsofpathogenesis
AT zhangpeilin thegeneexpressionprofilesofinducedpluripotentstemcellsfromindividualswithchildhoodcerebraladrenoleukodystrophyareconsistentwithproposedmechanismsofpathogenesis
AT luwange thegeneexpressionprofilesofinducedpluripotentstemcellsfromindividualswithchildhoodcerebraladrenoleukodystrophyareconsistentwithproposedmechanismsofpathogenesis
AT dranchakpatriciak thegeneexpressionprofilesofinducedpluripotentstemcellsfromindividualswithchildhoodcerebraladrenoleukodystrophyareconsistentwithproposedmechanismsofpathogenesis
AT shibatadarryl thegeneexpressionprofilesofinducedpluripotentstemcellsfromindividualswithchildhoodcerebraladrenoleukodystrophyareconsistentwithproposedmechanismsofpathogenesis
AT steinbergstevenj thegeneexpressionprofilesofinducedpluripotentstemcellsfromindividualswithchildhoodcerebraladrenoleukodystrophyareconsistentwithproposedmechanismsofpathogenesis
AT haciajosephg thegeneexpressionprofilesofinducedpluripotentstemcellsfromindividualswithchildhoodcerebraladrenoleukodystrophyareconsistentwithproposedmechanismsofpathogenesis
AT wangxiaoming geneexpressionprofilesofinducedpluripotentstemcellsfromindividualswithchildhoodcerebraladrenoleukodystrophyareconsistentwithproposedmechanismsofpathogenesis
AT yikwingyan geneexpressionprofilesofinducedpluripotentstemcellsfromindividualswithchildhoodcerebraladrenoleukodystrophyareconsistentwithproposedmechanismsofpathogenesis
AT zhangpeilin geneexpressionprofilesofinducedpluripotentstemcellsfromindividualswithchildhoodcerebraladrenoleukodystrophyareconsistentwithproposedmechanismsofpathogenesis
AT luwange geneexpressionprofilesofinducedpluripotentstemcellsfromindividualswithchildhoodcerebraladrenoleukodystrophyareconsistentwithproposedmechanismsofpathogenesis
AT dranchakpatriciak geneexpressionprofilesofinducedpluripotentstemcellsfromindividualswithchildhoodcerebraladrenoleukodystrophyareconsistentwithproposedmechanismsofpathogenesis
AT shibatadarryl geneexpressionprofilesofinducedpluripotentstemcellsfromindividualswithchildhoodcerebraladrenoleukodystrophyareconsistentwithproposedmechanismsofpathogenesis
AT steinbergstevenj geneexpressionprofilesofinducedpluripotentstemcellsfromindividualswithchildhoodcerebraladrenoleukodystrophyareconsistentwithproposedmechanismsofpathogenesis
AT haciajosephg geneexpressionprofilesofinducedpluripotentstemcellsfromindividualswithchildhoodcerebraladrenoleukodystrophyareconsistentwithproposedmechanismsofpathogenesis